基于流感嗜血杆菌外膜蛋白ompP6基因的PCR的建立和临床应用  

The establishment and clinical application of PCR based on the outer membrane protein om P6 gene of H. influenzae

作　　者：陆琴 金秀萍 汪琴琴 刘小香[3] 孙爱华[3] 

机构地区：[1]绍兴市柯桥区中医医院检验科,浙江绍兴312030 [2]绍兴市柯桥区中医医院儿科,浙江绍兴312030 [3]浙江医学高等专科学校基础医学部,浙江杭州310053

出　　处：《中国卫生检验杂志》2015年第4期457-459,共3页Chinese Journal of Health Laboratory Technology

基　　金：国家自然科学基金项目(81271893);浙江省自然科学基金项目(LY12H19002);浙江省医药卫生科技计划项目(2011KYA005)

摘　　要：目的建立基于流感嗜血杆菌外膜蛋白omp P6基因的PCR检测方法,并确定其检测限、灵敏度和特异性。方法根据Gen Bank公布的omp P6基因核苷酸序列设计引物,用流感嗜血杆菌标准菌株验证其检测限,通过检测1 225例呼吸道感染标本并与细菌培养法比较确定所建立的PCR检测方法的灵敏度与特异性。结果 1 225例呼吸道感染标本,细菌培养流感嗜血杆菌的阳性率为10.94%(134/1 225),PCR检测流感嗜血杆菌的阳性率为19.18%(235/1 225),培养法检测阳性标本PCR法检测均阳性。建立的PCR方法检测临床标本流感嗜血杆菌检出率明显高于细菌培养(χ2=32.5,P<0.01)。检测时模板的下限为3 pg,方法的灵敏度为100%,特异性为90.7%。结论建立的PCR方法具有快速和灵敏度高的特点,对于流感嗜血杆菌引起的呼吸道感染早期诊断有重要意义。Objective To establish a method of PCR based on the outer membrane protein om P6 gene of H. influenzae,and to evaluate its detection limit,sensitivity and specificity. Methods According to the omp P6 gene published Gen Bank to design primers,and use H. influenzae standard strains to verify the detection limit,and to further evaluate the sensitivity and specificity of the PCR based on culture using 1 225 respiratory infection specimens. Results In 1 225 specimens,positive rate of H. Influenzae were 10. 94%（134 /1 225） and 19. 18%（ 235 /1 225）,respestively based on culture and PCR. H. influenzae were positive specimens by culture,while they were also positive by PCR. The detection rate of H. Influenzae from clinical specimenswith PCR and significant higher than that with culture（ χ2= 32. 5,P 0. 01）. The lower limit of detection was 3 pg. The sensitivity and specificity of PCR were 100% and 90. 7%,respestively. Conclusion The established PCR is sensitive,specific,rapid and valuable for the early diagnosis of respiratory infection caused by H. influenzae.

关 键 词：流感嗜血杆菌 呼吸道感染 聚合酶连反应 

分 类 号：R378.4[医药卫生—病原生物学]